Method for producing 5'-flavour development nucleic acid
A nucleotide and nucleotide sequence technology, applied in the field of producing 5'-taste nucleotides, can solve the problems of low conversion rate and long production cycle, and achieve the effects of low cost, cost saving and short cycle
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0033] 1. Construction of genetically engineered bacteria
[0034] Primers were designed according to the phoc gene sequence of Enterobacter aerogenes provided in GeneBank as follows:
[0035] Primer1: 5'-cg ggatcc catgaaaaagcgcgttctcgccctctg-3'(BamHI) (denoted as SEQ.ID.NO.5),
[0036] Primer2: 5'-ccg ctcgag cgatgacgttacttctgcgttttggcg-3' (XholI) (denoted as SEQ.ID.NO.6),
[0037] Perform PCR using the chromosomal DNA of Enterobacter aerogenes as a template: 95°C for 5 minutes, 30× (95°C for 30s, 55°C for 30s, 72°C for 60s), 72°C for 10 minutes. A PCR product with a length of about 760bp was obtained, which was ligated with the pBV220 vector with T4 DNA ligase after restriction enzyme digestion, and transformed into E.coliDH5α competent cells. The pBV220 vector general primer PCR and restriction enzyme digestion were used to identify the transformants to obtain the genetically engineered strain pBV220-phoc. The DNA sequencing result is SEQ.ID.NO.1:
[0038]atgaaaaagcgc...
Embodiment 2
[0055] 1. Mutation of phoc
[0056] Design the following primers:
[0057] Primer3: 5'-ctaacctcagcttcggcgacgtggc-3' (denoted as SEQ.ID.NO.7),
[0058] Primer4: 5'-gccacgtcgccgaagctgaggttag-3' (denoted as SEQ.ID.NO.8),
[0059] Primer5: 5'-gcatacctctaccggctgggccac-3' (denoted as SEQ.ID.NO.9),
[0060] Primer6: 5'-gtggcccagccggtagagaggtatgc-3' (denoted as SEQ.ID.NO.10),
[0061] Using primer1 / primer4, primer3 / primer6, primer5 / primer2 as primers, pBV220-phoc recombinant plasmid as template, PCR, the conditions are as follows: 95°C for 5min, 30×(95°C for 30s, 50°C for 30s, 72°C for 60s), 72°C for 10 minutes. The PCR products with a length of about 200bp, 200bp, and 300bp were respectively obtained, and after recovery, they were used as templates and primer1 / primer2 were used as primers for the second round of PCR. ℃ 60s), 72 ℃ 10min. A PCR product with a length of about 760bp was obtained, which was ligated with the pBV220 vector with T4 DNA ligase after restriction enzyme d...
PUM

Abstract
Description
Claims
Application Information

- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com